Saudi Journal of Biological Sciences 29 (2022) 453–459

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Original article

Analysis of OCT1, OCT2 and OCT3 gene polymorphisms among Type 2

diabetes mellitus subjects in Indian ethnicity, Malaysia
Sabah Ghasan Abood Al-Ashoor a, Vasudevan Ramachandran b,1,⇑, Liyana Najwa Inche Mat a,d,
Nur Afiqah Mohamad a, Mohd Hazmi Mohamed c,d, Wan Aliaa Wan Sulaiman a,b,d,1,⇑
a
Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor DE, Malaysia
b
Centre for Materials Engineering and Regenerative Medicine, Bharath Institute of Higher Education and Research, 173, Agaram Main Rd, Selaiyur, Chennai, Tamil Nadu 600073, India
c
Department of Otorhinolaryngology-Head and Neck Surgery, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor DE, Malaysia
d
Malaysian Research Institute on Ageing, Universiti Putra Malaysia, Serdang 43400, Selangor DE, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Background: Type 2 Diabetes mellitus (T2DM) is a chronic metabolic disorder. It is a major non-
Received 15 April 2021 communicable disease affecting 463 million people globally in 2019 and is expected to be double to
Revised 5 September 2021 about 700 million by 2045. The majority are Asians with Indian ethnicity in Malaysia reported as the
Accepted 5 September 2021
highest prevalence of T2DM. Cardiovascular disease, renal failure, blindness and neuropathy, as well as
Available online 14 September 2021
premature death are the known morbidity and mortality resulted from T2DM. T2DM is characterized
by the dysfunctional insulin physiology that causes reduction of glucose transport into the cells which
Keywords:
lead to hyperglycaemia. Hence, one of the important treatments is an oral antidiabetic drug that lowers
Type 2 diabetes mellitus
Organic cation transporter
the serum glucose level in patients with T2DM. This drug will be transported across cell membranes by
OCT polymorphisms organic cation transporters (OCT). Therefore, it is important to identify the OCT candidate gene polymor-
Indian ethnicity phisms related to T2DM especially among the Indian ethnicity in Malaysia.
Methods: Blood samples were collected from 132 T2DM patients and 133 controls. Genotyping of OCT1
(rs628031), OCT2 (rs145450955), OCT3 (rs3088442 and rs2292334) was performed using (PCR-RFLP).
Results: No association was observed for genotypic and allelic distributions in all the gene polymor-
phisms of OCT genes (P > 0.05). However, a logistic regression analysis stratified by gender in a dominant
model showed a significant difference for OCT3 among males with T2DM (P = 0.006). Significant associ-
ation was also observed for OCT3 when stratified to subjects aged > 45 years old (P = 0.009).
Conclusion: Based on these findings, the association of OCT3 (rs2292334) could be considered as a pos-
sible genetic risk factor for the development of T2DM among Indian males alone.
Ó 2021 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access
article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction bolism caused by the body’s impaired ability to produce or respond

Diabetes mellitus is defined as ‘‘a chronic metabolic disease
characterized by hyperglycemia and abnormal carbohydrate meta-
⇑ Corresponding authors at: Department of Medicine, Faculty of Medicine and
Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor DE, Malaysia.
E-mail addresses: asuphd@gmail.com (S. Ghasan Abood Al-Ashoor), vasudevan-
ramachandran@bharathuniv.ac.in (V. Ramachandran), wanaliaa@upm.edu.my
(W.A. Wan Sulaiman).
1
These authors shared equal correspondence.
Peer review under responsibility of King Saud University.
Production and hosting by Elsevier
to insulin” (World Health Organization, 2019). Diabetes mellitus
can be classified into two classes, Type 1 diabetes (T1DM), which
is caused by deficit in total insulin secretion and Type 2 diabetes
mellitus (T2DM), a combination of inadequate insulin secretion
and resistance to insulin (Tuomi et al., 2014; Zolk, 2012).
Diabetes is developing rapidly around the world as the standard
of living rises (Wild et al., 2004). Around 415 million people world-
wide are believed to have diabetes, which is expected to reach 642
million by 2040 (Ogurtsova et al., 2017). Malaysia, a country com-
prising a multi-ethnic population, has one of the highest preva-
lence of T2DM compared to the other Asian countries, and the
trend is projected to continue to rise (Abdullah et al., 2017). The
National Health and Morbidity Surveys (NHMS) 2015, reported
the prevalence of T2DM in Malaysia to be highest among Indians
(22.1%), followed by Malays (14.6%), Chinese (12.0%), and indige-

https://doi.org/10.1016/j.sjbs.2021.09.008
1319-562X/Ó 2021 The Author(s). Published by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
S. Ghasan Abood Al-Ashoor, V. Ramachandran, Liyana Najwa Inche Mat et al.
nous Malaysian ethnicities (10.7%) (Institute for Public Health
NHMS, 2015).
T2DM is a chronic, progressive disorder that develops as a result
of the interaction between genetic and environmental factors
(overweight, obesity, high carbohydrate intake, and sedentary life-
style) (Chiasson and Rabasa-Lhoret, 2004). Treatments for diabetes
mellitus are aimed toward reducing microvascular and macrovas-
cular complications through optimal control (van Leeuwen et al.,
2013). Oral antidiabetic drugs are crucial as first-line treatments
for T2DM patients due to their mild hydrophobic property and
requirement of transporters to cross the plasma membrane into
the cells (Umamaheswaran et al., 2011).
A family of transporters known as ‘‘organic cation transporters”
(OCT1, OCT2, and OCT3) is involved in the transfer of endogenous
physiological amino compounds and is encoded by the Solute car-
rier family 22 member 1 SLC22A1, Solute carrier family 22 member
2 SLC22A2, and Solute carrier family 22 member 3 SLC22A3 genes,
respectively (Ghaffari-Cherati et al., 2016). The OCT (SLC22A) gene
family encodes>30 proteins and are mapped onto chromosome
6q25.3. The gene contains 11 exons and is involved in the excretion
of a wide variety of drugs, toxins, and metabolites in the kidney,
liver, and other tissues. Therefore, human reaction to anti-
diabetics medication is possibly influenced by genetic variants in
the OCT (SLC22A) genes (Hakooz et al., 2017; Kashi et al., 2015;
Kang et al., 2007).
The OCT family in humans consists of three organic cation
transporters (OCT1, OCT2, and OCT3), with OCT3 less studied com-
pared to OCT1 and OCT2 (Chen et al., 2015; Koepsell, 2004). It is
well established that OCT3 interferes with substrate specificity of
two other OCT family transporters and that it is an efficient trans-
porter for drugs (Hosseyni-Talei et al., 2017; Chen et al., 2015; Zhu
et al., 2012). OCT3, being a major transporter, also contributes to
the overall regulation of neurotransmission and the maintenance
of homeostasis within the central nervous system (Massmann
et al., 2014). OCT1 and OCT2 are expressed at different levels in
the liver, small intestine, kidney and brain (Koepsell, 2011). OCT1
is mainly expressed in the basolateral membrane of hepatocytes
that initiate uptake from the blood, whereas OCT2 is primarily
expressed in the basolateral membrane of kidney epithelial cells
(König et al., 2011; Meyer zu Schwabedissen et al., 2010; Müller
et al., 2013; Sato et al., 2008). Several factors including the nature
of the disease and the target organ, influence drug response; how-
ever, estimates indicate that genetic variation accounts for 20% to
95% of the variability in the response to similar anti-diabetic drugs
(Ghaffari-Cherati et al., 2016).
Several single nucleotide polymorphisms (SNPs) in the three
genes have been identified through population genetic analyses
studies (Du Plessis et al., 2015). These polymorphisms may con-
tribute to the inter-individual variability in the pharmacokinetics
disposition, efficacy, and toxicity of therapeutic drugs. Recent
researches have demonstrated inter-patient differences in
response to pharmacological therapy may be related to a variety
of genetic variants in the OCT genes (Du Plessis et al., 2015).
Early detection of high-risk individuals is critical for preventing
or delaying the onset of diabetes. Individuals develop diabetes as a
result of the interaction of genetic and environmental factors. Thus,
a susceptibility gene may exhibit a variable phenotype across pop-
ulations or regions as numerous studies have also shown inconsis-
tent findings regarding the association between OCT1, OCT2, and
OCT3 gene variants and diabetes in different ethnic groups
(Phate et al., 2020). There is a paucity of knowledge in Malaysia
about the OCT gene polymorphisms associated with T2DM. In
Malaysia, the Indian ethnic group has the greatest prevalence of
T2DM when compared to other ethnic groups (Institute for
Public Health NHMS, 2015). Thus, this study will ascertain the
genetic variation of the OCT1, OCT2, and OCT3 genes and their
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Saudi Journal of Biological Sciences 29 (2022) 453–459
association with T2DM in Malaysia’s Indian ethnic group. Addition-
ally, this study enhances our understanding of additional factors,
such as phenotypic characteristics, that might contribute to
T2DM and may have a significant impact on future individual med-
ical treatment for T2DM.
2. Materials and methods
2.1. Ethics statement
The present case-control study was registered with the Malay-
sian National Medical Research Register (NMRR) (reference num-
ber NMRR-18–3028-44490) and the study protocol was approved
by the Medical Research and Ethics Committee (MREC), Ministry
of Health (MoH), Malaysia.
2.2. Study population
All subjects were recruited from Hospital Serdang, Malaysia,
and were between the age of 18 and 65 years old and of Indian eth-
nicity. All participants were required to sign a written informed
consent form prior to participation in the study, and socio-
demographic profiles were obtained. Participants diagnosed with
T2DM by the medical practitioner were recruited as case subjects.
The T2DM diagnosis was based on a fasting plasma glucose (FPG)
of
7.0 mmol/L and HbA1c
6.3% following the T2DM clinical
practice guidelines from the Ministry of Health, Malaysia
(Ministry of Health, 2020). Healthy participants without T2DM
and any history of chronic or acute diseases (coronary artery,
myocardial infarction, stroke, T1DM, prior history of renal failure,
autoimmune disease, liver disease) were recruited as control sub-
jects. An amount of 2 to 3 ml of blood was collected into EDTA
tubes (Becton Dickinson, New Jersey, USA) from all subjects and
stored at 20 °C for further analysis.
2.3. Gene and SNP selection
Based on the obtained literature reviews, OCT1 (rs628031),
OCT2 (rs145450955), and OCT3 (rs3088442 and rs2292334) gene
polymorphisms were among the significant SNPs studied in
T2DM (Mahrooz et al., 2017), and thus, were selected as a candi-
date SNP in the present study. Furthermore, the SNP was selected
based on functional significance and location. All the SNPs were
not in linkage disequilibrium, which were performed using the
Haploview software (http://www.broad.mit.edu/mpg/haploview).
The RegulomeDB (http://www.regulomedb.org/) was used to
determine the effect of the OCT SNPs on allele-specific transcrip-
tion factor binding and link on the expression of the gene target.
All SNPs scored a RegulomeDB of 4 and 5 which indicate the vari-
ants had minimal transcription factor binding evidence.
2.4. Genotyping
Genomic DNA was extracted using the QIAamp DNA blood mini
kit (Qiagen, Valencia, CA, USA) following the manufacturer’s proto-
col. Genotyping was then performed using PCR-RFLP method for
the four gene polymorphisms; OCT1 (rs628031), OCT2
(rs145450955), and OCT3 (rs3088442 and rs2292334). The PCR
was carried out in a 25 mL volume of a reaction mixture comprising
1 unit of Taq DNA polymerase, 400 to 500 ng genomic DNA,
200 mM of each dNTP, 1.5 mM MgCL2, 280 nM of the following
primers: forward: 50 -CTAAACCCAGTGATTCATGCTCTTT-3 ’ and
reverse: 50 -TTTGTTCTCATT-CCAGAGGCTTATC-30 for OCT1
(rs628031); forward: 50 -CTGTGCCTCCTGGATTCATA-30 and reverse:
50 -AGGTTGCTTTTGTTCTACAGT-30 for OCT2 (rs145450955);
S. Ghasan Abood Al-Ashoor, V. Ramachandran, Liyana Najwa Inche Mat et al.
forward: 50 -AGATTGCATGGAGGATGAAC-30 and reverse: 50 -TGT
TCCAGAGGAGGTGGACG-30 for OCT3 (rs3088442); forward: 50 -GT
GGTGGAACTGCCAGGA-30 and reverse: 50 -CTACAGAACCAATCTCT
TACTTCG-30 for OCT3 (rs2292334). The PCR cycling conditions were
carried out for 35 cycles with denaturation at 94 °C for 30 s
(rs628031), 93 °C for 35 s (rs145450955), 93 °C for 50 s
(rs3088442 and rs2292334), annealing at 56 °C for 60 s
(rs628031), 54 °C for 35 s (rs145450955), 54 °C for 40 s
(rs3088442), 55 °C for 40 s (rs2292334), and extended at 72 °C
for 60 s (rs628031), 35 s (rs145450955) and 40 s (rs3088442 and
rs2292334) respectively. The amplification products were digested
with MscI (rs628031), BsaAI (rs145450955) (Fermentas, Lithuania),
and AciI (rs3088442 and rs2292334) restriction enzymes (Thermo
Scientific, Lithuania) and visualized under UV light with the Alpha
Imager (Alpha Innotech, San Leandro, CA).
2.5. Statistical analysis
All statistical data were analysed using IBM SPSS software (ver-
sion 24.0, SPSS Inc., Chicago, USA). The Hardy-Weinberg equilib-
rium and linkage disequilibrium of alleles in each SNPs were
confirmed using the Haploview software (http://www.broad.mit.
edu/mpg/haploview). Categorical variables were compared using
the chi-square test, while normal continuous variables using the
Student’s t-test and one-way analysis of variance (ANOVA).
Skewed continuous variables are compared using Mann-Whitney
U test. Logistic regression analysis was performed to determine
the association between OCT1 (rs628031), OCT2 (rs145450955),
and OCT3 (rs3088442 and rs2292334) gene polymorphisms with
T2DM under the assumption of three genetic models (additive,
dominant and recessive) and also stratified by gender. Bonferroni
adjustment for multiple statistical testing was applied to correct
the threshold of P-value. Any subjects with missing data were
excluded from the study and a P-value of p < 0.05 was considered
statistically significant.
3. Results
3.1. Clinical and biochemical parameters of study subjects
A total of 132 T2DM patients and 133 controls were recruited
based on the inclusion and exclusion criteria. Table 1 illustrates
the clinical and parameters between T2DM and control subjects.
T2DM subjects have a mean age of 49.83 ± 13.68 years, whereas
the control subjects with a mean of 46.65 ± 14.20 years. There is
a significant difference in HbA1c levels between T2DM and con-
trols (P < 0.001), with higher level of HbA1c among the control sub-
jects (8.26 ± 1.89).
3.2. Genotypic and allelic distributions
Genotyping was performed for all the four OCT gene polymor-
phisms as presented in Fig. 1. No significant association was
recorded in both genotype and allele frequency in the four OCT
gene polymorphisms when compared between T2DM and control
subjects (P > 0.05). A significant association (P = 0.009) was
observed only for OCT3 (rs2292334) gene polymorphism when
stratified to subjects aged > 45 years old (Table 2). Likewise, all
gene variants when compared based on an additive, dominant
and recessive genetic model were not significantly different
(P > 0.05) (Table 3). However, a significant difference was observed
in OCT3 (rs2292334) gene polymorphism in a dominant genetic
model when stratified by male gender and for all ages
(P = 0.006). No significant difference was recorded for the other
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Saudi Journal of Biological Sciences 29 (2022) 453–459
gene polymorphisms after stratifying by gender (P > 0.05)
(Table 4).
4. Discussion
A total of 265 subjects were recruited for this study, including
132 subjects with T2DM and 133 control subjects. To our knowl-
edge, this is the first study reporting the frequency of OCT1,
OCT2 and OCT3 gene polymorphisms among Malaysia’s Indian eth-
nic group. From our findings, there was no significant association
in both genotypic and allelic distribution observed in OCT1
(rs628031), OCT2 (rs145450955), OCT3 (rs3088442 and
rs2292334) gene polymorphisms between T2DM and control sub-
jects (P > 0.05). The allelic distributions of all OCT gene polymor-
phisms studied on T2DM in different populations are shown in
Table 5 with contradictory findings. These inconsistent results
could be explained by various factors including ethnic differences,
environmental factors, and different study methodology.
Apart from the genetic factors, clinical and biochemical param-
eters such as abdominal obesity, family history of diabetes and
hypertension, older age and poor diet are associated with diabetes.
Recently, the Malaysian Ministry of Health projected that the
prevalence of T2DM could rise to as much as 20.1% in 2020 if the
major associated factors (overweight/obesity, poor diet and inade-
quate physical activity) are not comprehensively addressed
(Ministry of Health, 2013; Institute for Public Health NHMS,
2015). The average age of T2DM onset is 45 years old and a 10-
year prospective cohort study conducted in India reported that
subjects older than 45 years had a1.6 times higher risk of develop-
ing T2DM (Vijayakumar et al., 2019). Hence, the present study
included an additional analysis excluding those<45 years old as
there might be a possibility of the young subjects underpowering
the study since they might be future cases carrying the genetic
variants.
In this study, a significant association was recorded for the
rs2292334 variant of the OCT3 gene when analyzed among sub-
jects aged>45 years old (P = 0.009). A similar variant also showed
a significant difference when stratified by the male gender
(P < 0.05). The mutant variant was also seen to be at the highest
risk compared to the other variants.
A study by Chen et al. reported that rs2292334 is involved in the
therapeutic action of metformin and with an A allele frequency of
0.45 could be considered as a polymorphic variant in the popula-
tion even with different ethnic groups (Chen et al., 2010). This is
consistent with the present study and also a study among the Ira-
nian population that reported a frequency of 0.65 major G alleles
and 0.35 of minor A alleles of the rs2292334 variant in T2DM
patients (Mahrooz et al., 2017).
According to Aoyama et al., (2006) and Nies et al., (2009), the
rs2292334 variant plays a critical role in the gene expression of
OCT3 and influences the final production of mRNA. The minor A
allele of rs2292334 was also linked to a higher risk of T2DM among
obese patients in comparison with non-obese patients (Mahrooz
et al., 2017). A combination of obesity and parental history of dia-
betes among males has also shown a significant interaction that
increases the risk of T2DM (OR = 2.4) (Wikner et al., 2013). This fur-
ther proves the findings from the present study that showed a sig-
nificant association of OCT3 rs2292334 gene polymorphism with
Indian ethnicity male subjects with T2DM. In the Pakistani popula-
tion, the A allele of OCT3 rs3088442 was a protective factor and
associated with the clinical efficacy of metformin in patients who
have type 2 diabetes (Moeez et al., 2019).
Despite the findings, the present study has several limitations.
This study lacked access to data on gene expression, which could
be used to assess the association between corresponding polymor-
S. Ghasan Abood Al-Ashoor, V. Ramachandran, Liyana Najwa Inche Mat et al. Saudi Journal of Biological Sciences 29 (2022) 453–459

Table 1
Clinical and biochemical parameters of study subjects.

Parameters T2DM (n = 132) Controls (n = 133) P-valuey

Age (Years) Total 49.83 ± 13.68 46.65 ± 14.20 0.069
45 years 47 (35.6) 61 (45.9)
>45 years 85 (64.4) 72 (54.1) 0.089§
Gender Male 84 (63.6) 71 (53.4)
Female 48 (36.4) 62 (46.6) 0.09§
BMI (Kg/m2) 27.83 ± 4.72 27.70 ± 4.96 0.625à
Blood glucose FBS(mmol/L) 7.84 ± 3.00 7.47 ± 2.64 0.669
HbA1C 7.84 ± 6.61 8.26 ± 1.89 <0.001*

Data are presented as mean ± SD and data reported as number (percentage) of subjects for gender; BMI = body mass index, FBS = fasting blood sugar, HbA1C = glycated
haemoglobin
*Significant P-value, P < 0.05
y
Mann-Whitney U test
à
Student’s t-test
§
Pearson chi-square test

Table 2
Genotypic and allelic distributions of SLC22A1 rs628031, SLC22A2 rs145450955, SLC22A3 rs3088442 and SLC22A3 rs2292334 gene polymorphisms between T2DM and control
subjects.

SNPs Genotypes (%) P-valuey Alleles (%) P-valuey Odd ratio

(95% CI)
All age
rs628031 T2DM AA AG GG A G
Controls 70(53.4) 51(38.9) 10(7.7) 0.336 191(72.9) 71(27.1) 0.157 1.333
83(62.4) 42(31.6) 8 (6.0) 208(78.2) 58(21.8) (0.895–1.986)
rs145450955 T2DM CC CT TT C T
Controls 71(53.8) 17(12.9) 44(33.3) 0.431 159(60.2) 105(39.8) 0.11 1.336
82(61.7) 14(10.5) 37(27.8) 178(66.9) 88(33.1) (0.937–1.905)
rs3088442 GG GA AA G A
T2DM 83(62.9) 16(12.1) 33(25.0) 0.89 182(68.9) 82(31.1) 0.531 0.888
Controls 87(65.4) 16(12.0) 30(22.6) 190(31.1) 76(28.6) (0.612–1.288)
rs2292334 GG GA AA G A
T2DM 82(62.1) 19(14.4) 31(23.5) 0.29 183(69.3) 81(30.7) 0.05 1.473
Controls 94(71.2) 15(11.4) 23(17.4) 203(76.9) 61(23.1) (1.000–2.171)
Age > 45 years
rs628031 T2DM AA AG GG A G
Controls 44(50.6) 34(56.7) 7(70.0) 0.446 122(71.8) 48(28.2) 0.223 1.377
43(49.4) 26(43.3) 3(30.0) 112(78.9) 32(22.2) (0.822–2.306)
rs145450955 T2DM CC CT TT C T
Controls 43(51.2) 12(60.0) 30(56.6) 0.705 98(57.6) 72(42.4) 0.382 1.224
41(48.8) 8(40.0) 23(43.4) 90(62.5) 54(37.5) (0.777–1.929)
rs3088442 GG GA AA G A
T2DM 49(50.0) 12(60.0) 24(61.5) 0.404 110(64.7) 60(35.3) 0.090 1.522
Controls 49(50.0) 8(40.0) 15(38.5) 106(73.6) 38(26.4) (0.936–2.474)
rs2292334 GG GA AA G A
T2DM 52(49.1) 10(62.5) 23(67.6) 0.132 114(67.1) 56(32.9) 0.009* 2.000
Controls 54(50.9) 6(37.5) 11(32.4) 114(79.2) 28(19.4) (1.186–3.373)

SNPs = single nucleotide polymorphisms, T2DM = type 2 diabetes mellitus, CI = confidence interval.
*Significant P-value, P < 0.05.
y
Pearson Chi-square test.

Table 3
Genotypes association based on genetic models.

OR (95% CI) for diabetics

Gene variant Additive Dominant Recessive py pà p§
All age
rs628031 1.49 (0.55–4.06) 1.37 (0.84–2.26) 1.34 (0.50–3.56) 0.428 0.212 0.56
rs145450955 1.29 (0.75–2.24) 1.28 (0.78–2.11) 1.25 (0.73–2.12) 0.359 0.335 0.419
rs3088442 1.09 (0.61–1.96) 1.11 (0.67–1.85) 1.07 (0.60–1.89) 0.772 0.685 0.824
rs2292334 1.56 (0.84–2.91) 1.61 (0.95–2.73) 1.43 (0.78–2.64) 0.163 0.075 0.249
Age > 45 years
rs628031 2.26 (0.55–9.34) 1.39 (0.73–2.62) 2.05 (0.51–8.23) 0.258 0.313 0.314
rs145450955 1.27 (0.63–2.53) 1.31 (0.69–2.47) 1.18 (0.61–2.31) 0.506 0.404 0.625
rs3088442 1.56 (0.72–3.35) 1.53 (0.79–2.97) 1.45 (0.69–3.07) 0.259 0.207 0.328
rs2292334 2.17 (0.96–4.90) 2.04 (1.01–4.10) 2.02 (0.90–4.49) 0.062 0.046 0.087

OR = odds ratio, CI = confidence interval.

py, for the additive model; pà, for the dominant model; and p§, for the recessive model.
yà§
Logistic regression analysis

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S. Ghasan Abood Al-Ashoor, V. Ramachandran, Liyana Najwa Inche Mat et al. Saudi Journal of Biological Sciences 29 (2022) 453–459

Table 4
Genotypes associations based on genetic models and genders.

OR (95% CI)
Age Gender Gene Variant Additive Dominant Rececssive py pà p§
All Male rs628031 1.97 (0.50–7.71) 1.77 (0.90–3.48) 1.37 (0.37–5.06) 0.332 0.756 0.633
rs145450955 1.03 (0.49–2.18) 1.05 (0.54–2.04) 1.12 (0.56–2.22) 0.937 0.885 0.755
rs3088442 0.81 (0.37–1.77) 0.81 (0.40–1.66) 0.94 (0.45–1.96) 0.6 0.569 0.866
rs2292334 3.11 (1.25–7.73) 2.91 (1.37–6.19) 2.92 (1.21–7.08) 0.014 0.006* 0.018
Female rs628031 0.69 (0.11–4.11) 0.83 (0.36–1.89) 1.09 (0.21–5.67) 0.679 0.658 0.917
rs145450955 1.78 (0.70–4.51) 1.99 (0.86–4.63) 1.56 (0.63–3.84) 0.224 0.107 0.338
rs3088442 1.57 (0.54–4.55) 1.43 (0.63–3.22) 1.22 (0.45–3.31) 0.407 0.392 0.703
rs2292334 0.53 (0.18–1.52) 0.70 (0.31–1.61) 0.55 (0.19–1.50) 0.237 0.401 0.241
>45 years Male rs628031 1.45 (0.29–7.13) 1.91 (0.84–4.31) 1.07 (0.23–5.07) 0.651 0.121 0.930
rs145450955 1.13 (0.45–2.83) 1.09 (0.49–2.48) 1.11 (0.47–2.63) 0.799 0.826 0.808
rs3088442 1.69 (0.66–4.29) 1.82 (0.76–4.33) 1.55 (0.62–3.88) 0.273 0.178 0.353
rs2292334 3.30 (1.09–10.1) 2.69 (1.04–6.93) 3.14 (1.04–9.47) 0.035 0.040 0.042
Female rs628031 NA 0.73 (0.25–2.12) NA NA 0.565 NA
rs145450955 1.28 (0.43–3.86) 1.49 (0.51–4.41) 1.15 (0.39–3.45) 0.659 0.462 0.797
rs3088442 1.25 (0.32–4.91) 1.22 (0.42–3.50) 1.19 (0.32–4.45) 0.748 0.713 0.795
rs2292334 1.23 (0.34–4.37) 1.43 (0.48–4.19) 1.11 (0.32–3.84) 0.752 0.520 0.872

OR = odds ratio, CI = confidence interval, NA = not available.

py, for the additive model; pà, for the dominant model; and p§, for the recessive model.
*Significant P-value, P < 0.01 after Bonferroni correction (0.05/4 for 4 variants).
yà§
Logistic regression analysis, adjusted for age.

Table 5
Allele frequency distribution of OCT gene polymorphisms.

SNPs Population Case/control Alleles in cases Alleles in controls P-value Reference

G A G A
OCT1 China 153/124 0.72 0.28 0.74 0.26 * Zhou et al., 2015
rs628031 Indonesia 86/NA 0.61 0.39 NA NA NS Vitarani et al., 2017
Saudi Arabia 201/89 0.69 0.31 0.71 0.29 NS Altall et al., 2019
Latvia 53/193 0.27 0.73 0.24 0.76 * Tarasova et al., 2012
Iran 63/77 0.32 0.68 0.33 0.66 NS Shokri et al., 2016
Present study 132/133 0.27 0.73 0.22 0.78 NS
C T C T
OCT2 Iran 40/NA 0.8 0.2 NA NA * Kashi et al., 2015
rs145450955 Present study 132/133 0.6 0.4 0.67 0.33 NS
G A G A
OCT3 Iran 150/152 NR NR NR NR ** Mahrooz et al., 2017
rs3088442 Pakistan 600/300 0.67 0.33 0.54 0.46 ** Moeez et al., 2019
Present study 132/133 0.69 0.31 0.31 0.29 NS
G A G A
OCT3 Iran 150/152 0.65 0.35 0.75 0.25 * Mahrooz et al., 2017
rs2292334 Present study 132/133 0.69 0.31 0.77 0.23 NS

NA, not available; NR, not reported; NS, not significant; *(p < 0.05); **(P < 0.001).

phisms. Additionally, this study’s findings are confined among selected based on their low minor allele frequency and potential
Malaysian Indians. Future research should include Malays and Chi- significance. To our best knowledge, this study is the first to eval-
nese as well, to better understand the genetic risk factor for OCT uate the association of OCT gene polymorphisms with T2DM
gene variants to develop in T2DM patients in a general Malaysian among the Indian ethnicity in Malaysia.
population. While advanced age is a risk factor for T2DM, this
study enrolled patients of adults aged above 18 years old. How-
5. Conclusion
ever, the current study includes a stratified analysis of participants
aged over 45 years to address this limitation. The contradictory
In conclusion, the association of OCT3 rs2292334 gene poly-
findings from this study in contrast to other populations could be
morphism could be considered as a genetic susceptibility to the
attributed to a variety of reasons, including ethnic diversity which
development of T2DM among Malaysian Indian ethnicity male
can result in distinctly different genetic predispositions. Certain
subjects. However, further study with a larger sample size is
countries may have heterogeneous ethnicities; this may result in
needed to confirm the association. This study could be used as a
different genetic predispositions in comparison to countries with
platform to further understanding genetic factors that cause
homogenous ethnicity. There is also the matter of varying sample
T2DM and, in the future, may have a significant impact on individ-
sizes in comparison with other epidemiological studies. In this
ualizing medical treatment for T2DM particularly for Indian
study, the relatively small number of samples and low frequency
ethnicity.
of the risk allele could contribute to the lack of association.
Although GWAS studies have suggested that OCT gene polymor-
phisms were associated with T2DM in various populations, there CRediT authorship contribution statement
is a lack of information on the relationship between OCT gene poly-
morphisms and Malaysian Indians Therefore, the candidate gene Sabah Ghasan Abood Al-Ashoor: Formal analysis, Investiga-
approach used in this study focuses on a few SNPs that were tion, Writing – original draft, Writing – review & editing. Vasude-
457
S. Ghasan Abood Al-Ashoor, V. Ramachandran, Liyana Najwa Inche Mat et al.
van Ramachandran: Conceptualization, Methodology, Validation,
Resources, Writing – review & editing, Supervision, Project admin-
istration, Funding acquisition. Liyana Najwa Inche Mat: Valida-
tion, Resources, Writing – review & editing, Supervision. Nur
Afiqah Mohamad: Validation, Formal analysis, Data curation,
Writing – review & editing. Mohd Hazmi Mohamed: Validation,
Resources, Writing – review & editing, Supervision. Wan Aliaa
Wan Sulaiman: Conceptualization, Methodology, Resources, Writ-
ing – review & editing, Supervision, Project administration, Fund-
ing acquisition.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
Acknowledgments
The authors would also like to put on record their appreciation
to the participants of the study for their cooperation and involve-
ment in this study.
Funding
This work was supported by the Putra Grant (Grant no: GP-
IPS/2018/9648800) from Universiti Putra Malaysia.
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